Production of Lactate by Metabolically Engineered <i>Scheffersomyces stipitis</i>
Lactate is a valuable compound used in food, chemical, and pharmaceutical industries. High-value, optically pure L- or D-lactate can be synthesized microbially via specific dehydrogenases. The non-conventional yeast <i>Scheffersomyces stipitis</i>, which is known for fermenting both hexo...
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2025-05-01
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| Online Access: | https://www.mdpi.com/2309-608X/11/6/413 |
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| author | Angela Matanović Nenad Marđetko Ana Slišković Blanka Didak Karla Hanousek Čiča Bojan Žunar Anamarija Štafa Božidar Šantek Marina Svetec Miklenić Ivan-Krešimir Svetec |
| author_facet | Angela Matanović Nenad Marđetko Ana Slišković Blanka Didak Karla Hanousek Čiča Bojan Žunar Anamarija Štafa Božidar Šantek Marina Svetec Miklenić Ivan-Krešimir Svetec |
| author_sort | Angela Matanović |
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| description | Lactate is a valuable compound used in food, chemical, and pharmaceutical industries. High-value, optically pure L- or D-lactate can be synthesized microbially via specific dehydrogenases. The non-conventional yeast <i>Scheffersomyces stipitis</i>, which is known for fermenting both hexoses and pentoses, is a promising host for biochemical production from lignocellulosic biomass but does not naturally produce lactate. In this study, we engineered <i>S. stipitis</i> to produce lactate by expressing two codon-optimized bacterial L-lactate dehydrogenase genes under the control of strong native promoters. The engineered strain produced 7.42 g/L (0.46 g/g yield) and 11.67 g/L (0.58 g/g yield) lactate from glucose and xylose, respectively. The highest titer, 19.27 g/L (0.52 g/g yield), was achieved from 50 g/L xylose after 74 h. Increasing the fermentation temperature from 28 °C to 32 °C improved yield by 30%, while a neutralizing agent further enhanced yield by 25% and prevented lactate degradation following carbon depletion. Although the wildtype strain produced a significant amount of ethanol on both glucose and xylose, the engineered strain produced ethanol as a side product exclusively on glucose and not on xylose. This phenomenon could be advantageous for biotechnological applications and may reflect shifts in gene expression depending on the carbon source or even on the presence of lactate. |
| format | Article |
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| institution | Kabale University |
| issn | 2309-608X |
| language | English |
| publishDate | 2025-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Journal of Fungi |
| spelling | doaj-art-47b54aa388404e358334dce036f0d56f2025-08-20T03:27:40ZengMDPI AGJournal of Fungi2309-608X2025-05-0111641310.3390/jof11060413Production of Lactate by Metabolically Engineered <i>Scheffersomyces stipitis</i>Angela Matanović0Nenad Marđetko1Ana Slišković2Blanka Didak3Karla Hanousek Čiča4Bojan Žunar5Anamarija Štafa6Božidar Šantek7Marina Svetec Miklenić8Ivan-Krešimir Svetec9Laboratory for Biology and Microbial Genetics, Department of Biochemical Engineering, Faculty of Food Technology and Biotechnology, University of Zagreb, Pierottijeva 6, 10000 Zagreb, CroatiaLaboratory for Biochemical Engineering, Industrial Microbiology and Malting and Brewing Technology, Department of Biochemical Engineering, Faculty of Food Technology and Biotechnology, University of Zagreb, Pierottijeva 6, 10000 Zagreb, CroatiaLaboratory for Biology and Microbial Genetics, Department of Biochemical Engineering, Faculty of Food Technology and Biotechnology, University of Zagreb, Pierottijeva 6, 10000 Zagreb, CroatiaLaboratory for Biochemical Engineering, Industrial Microbiology and Malting and Brewing Technology, Department of Biochemical Engineering, Faculty of Food Technology and Biotechnology, University of Zagreb, Pierottijeva 6, 10000 Zagreb, CroatiaLaboratory for Fermentation and Yeast Technology, Department of Food Engineering, Faculty of Food Technology and Biotechnology, University of Zagreb, Pierottijeva 6, 10000 Zagreb, CroatiaLaboratory for Biology and Microbial Genetics, Department of Biochemical Engineering, Faculty of Food Technology and Biotechnology, University of Zagreb, Pierottijeva 6, 10000 Zagreb, CroatiaLaboratory for Biology and Microbial Genetics, Department of Biochemical Engineering, Faculty of Food Technology and Biotechnology, University of Zagreb, Pierottijeva 6, 10000 Zagreb, CroatiaLaboratory for Biochemical Engineering, Industrial Microbiology and Malting and Brewing Technology, Department of Biochemical Engineering, Faculty of Food Technology and Biotechnology, University of Zagreb, Pierottijeva 6, 10000 Zagreb, CroatiaLaboratory for Biology and Microbial Genetics, Department of Biochemical Engineering, Faculty of Food Technology and Biotechnology, University of Zagreb, Pierottijeva 6, 10000 Zagreb, CroatiaLaboratory for Biology and Microbial Genetics, Department of Biochemical Engineering, Faculty of Food Technology and Biotechnology, University of Zagreb, Pierottijeva 6, 10000 Zagreb, CroatiaLactate is a valuable compound used in food, chemical, and pharmaceutical industries. High-value, optically pure L- or D-lactate can be synthesized microbially via specific dehydrogenases. The non-conventional yeast <i>Scheffersomyces stipitis</i>, which is known for fermenting both hexoses and pentoses, is a promising host for biochemical production from lignocellulosic biomass but does not naturally produce lactate. In this study, we engineered <i>S. stipitis</i> to produce lactate by expressing two codon-optimized bacterial L-lactate dehydrogenase genes under the control of strong native promoters. The engineered strain produced 7.42 g/L (0.46 g/g yield) and 11.67 g/L (0.58 g/g yield) lactate from glucose and xylose, respectively. The highest titer, 19.27 g/L (0.52 g/g yield), was achieved from 50 g/L xylose after 74 h. Increasing the fermentation temperature from 28 °C to 32 °C improved yield by 30%, while a neutralizing agent further enhanced yield by 25% and prevented lactate degradation following carbon depletion. Although the wildtype strain produced a significant amount of ethanol on both glucose and xylose, the engineered strain produced ethanol as a side product exclusively on glucose and not on xylose. This phenomenon could be advantageous for biotechnological applications and may reflect shifts in gene expression depending on the carbon source or even on the presence of lactate.https://www.mdpi.com/2309-608X/11/6/413microbial lactate production<i>Scheffersomyces stipitis</i>non-conventional yeastL-lactate dehydrogenasemetabolic engineeringxylose fermentation |
| spellingShingle | Angela Matanović Nenad Marđetko Ana Slišković Blanka Didak Karla Hanousek Čiča Bojan Žunar Anamarija Štafa Božidar Šantek Marina Svetec Miklenić Ivan-Krešimir Svetec Production of Lactate by Metabolically Engineered <i>Scheffersomyces stipitis</i> Journal of Fungi microbial lactate production <i>Scheffersomyces stipitis</i> non-conventional yeast L-lactate dehydrogenase metabolic engineering xylose fermentation |
| title | Production of Lactate by Metabolically Engineered <i>Scheffersomyces stipitis</i> |
| title_full | Production of Lactate by Metabolically Engineered <i>Scheffersomyces stipitis</i> |
| title_fullStr | Production of Lactate by Metabolically Engineered <i>Scheffersomyces stipitis</i> |
| title_full_unstemmed | Production of Lactate by Metabolically Engineered <i>Scheffersomyces stipitis</i> |
| title_short | Production of Lactate by Metabolically Engineered <i>Scheffersomyces stipitis</i> |
| title_sort | production of lactate by metabolically engineered i scheffersomyces stipitis i |
| topic | microbial lactate production <i>Scheffersomyces stipitis</i> non-conventional yeast L-lactate dehydrogenase metabolic engineering xylose fermentation |
| url | https://www.mdpi.com/2309-608X/11/6/413 |
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